Comprehensive diagnosis of Alzheimer's disease by detection of misfolded oligomers in biological fluids

通过检测生物体液中错误折叠的寡聚物来全面诊断阿尔茨海默病

• 批准号: 9766691
• 负责人: CLAUDIO SOTO
• 金额: $ 289.76万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9766691
• 关键词: Affect; Alzheimer disease detection; Alzheimer' s Disease; Amyloid beta-Protein; Amyotrophic Lateral Sclerosis; Appearance; Ataxia; Biochemical; Biological; Biological Assay; Blood; Brain; Brain Injuries; Cause of Death; Cerebrum; Clinical; DNA-Binding Proteins; Dementia; Deposition; Detection; Development; Diagnosis; Disease; Disease Progression; Early Diagnosis; Elderly; Encephalitis; Event; Fingerprint; Frontotemporal Dementia; Functional disorder; Goals; Human; Huntington Disease; Huntington gene; Individual; Inherited; Laboratory Diagnosis; Lead; Lewy Body Dementia; Liquid substance; Medical; Molecular Conformation; Monitor; Nerve Degeneration; Neurodegenerative Disorders; Neurofibrillary Tangles; Parkinson Disease; Parkinson' s Dementia; Pathogenesis; Pathologic; Pathology; Patients; Periodicity; Plasma; Plasma Proteins; Polymers; Population; PrP; Predictive Value; Prion Diseases; Process; Property; Proteins; Sampling; Seeds; Senile Plaques; Sensitivity and Specificity; Structure; Symptoms; Synapses; Tauopathies; Technology; Testing; Time; Urine; Vascular Dementia; alpha synuclein; amplification detection; clinical Diagnosis; disease diagnosis; extracellular; mild cognitive impairment; misfolded protein; neuron loss; noninvasive diagnosis; novel strategies; polymerization; pre-clinical; protein TDP-43; protein aggregate; protein aggregation; protein misfolding; protein misfolding cyclic amplification; response; synucleinopathy; tau Proteins; tau aggregation

ABSTRACT This is a revised application in response to the PAR-15-359, entitled “Novel Approaches to Diagnosing Alzheimer's Disease & Predicting Progression (R01)”. Alzheimer's disease (AD) is the most common dementia in the elderly population and one of the leading causes of death in the developed world. One of the main problems in AD is the lack of an early, sensitive and objective laboratory diagnosis to identify individuals that will develop the disease before substantial brain damage. Compelling evidences point that the hallmark event in AD is the misfolding, oligomerization and brain accumulation of protein aggregates. Similar to AD several other neurodegenerative disorders are associated to the accumulation of cerebral protein aggregates, including Parkinson disease (PD), amyotrophic lateral sclerosis, prion diseases, and Huntington disease. In AD the most abundant protein aggregates are amyloid plaques and neurofibrillary tangles composed of the amyloid-beta (Aβ) and Tau proteins, respectively. Although the protein involved in the misfolding and aggregation process is different in each disease, the end products and intermediate structures are very similar. Moreover, in all cases, protein misfolding and aggregation follows a seeding-nucleation mechanism in which the limiting step is the formation of small oligomeric intermediates that act as seeds to catalyze the polymerization process. Recent evidence has shown that misfolded oligomers are circulating in biological fluids and these structures appear to be key for inducing brain degeneration. Our working hypothesis is that a sensitive, comprehensive and early biochemical diagnosis of AD may be developed by ultra-sensitive and simultaneous detection of misfolded oligomers composed of the three most common protein aggregating in the human brain (Aβ, Tau, and αSyn) in human biological fluids (CSF and blood plasma). Our approach for detection of these oligomers is to use the functional property of misfolded oligomers of being capable to catalyze the polymerization of the monomeric protein as a way to detect them. For this purpose, we invented the protein misfolding cyclic amplification (PMCA), which represents a platform technology to detect very small quantities of seeding-competent misfolded oligomeric proteins associated with various protein misfolding diseases. Currently, PMCA has been adapted to detect misfolded prion protein implicated in prion diseases in various biological fluids, including blood and urine and more recently soluble Aβ and αSyn oligomers in CSF of AD and PD patients, respectively. The major goal of this project is to adapt the PMCA technology for specific and highly sensitive detection of misfolded Aβ, Tau, and αSyn oligomers in human blood and CSF in order to obtain a fingerprint profile that enable to diagnose AD and distinguish from other diseases with similar clinical presentation. In this project we will perform studies of specificity and sensitivity using large number of samples and evaluate the utility of PMCA for monitoring disease progression and for pre-clinical identification of people in the way to develop AD. The results generated in this project may lead to the development of a comprehensive biochemical test for AD that may be useful not only to aid in the diagnosis of the disease and differentiate it from other diseases with similar clinical presentation, but also to identify people on the way to developing AD pathology before the onset of substantial brain damage and clinical symptoms of the disease.

摘要 这是响应PAR-15-359的修订申请，标题为“ 诊断阿尔茨海默病和预测进展（R 01）。 阿尔茨海默病（AD）是老年人群中最常见的痴呆症，也是老年痴呆症的主要病因之一。 是发达国家的主要死因AD的主要问题之一是缺乏 早期，敏感和客观的实验室诊断，以确定个人将发展 在严重的脑损伤之前。令人信服的证据表明， AD是蛋白质聚集体的错误折叠、寡聚化和脑积聚。类似于AD 其他几种神经变性疾病与脑蛋白的积累有关 聚集体，包括帕金森病（PD）、肌萎缩性侧索硬化症、朊病毒疾病和 亨廷顿病。在AD中，最丰富的蛋白质聚集体是淀粉样斑块， 分别由β淀粉样蛋白（Aβ）和Tau蛋白组成的神经元缠结。 虽然在每个细胞中参与错误折叠和聚集过程的蛋白质是不同的， 疾病，最终产品和中间结构非常相似。此外，在所有情况下， 蛋白质的错误折叠和聚集遵循一种种子成核机制， 步骤是形成小的低聚中间体，其充当种子以催化聚合反应。 聚合过程最近的证据表明，错误折叠的低聚物在体内循环， 生物液体和这些结构似乎是诱发脑退化的关键。 我们的工作假设是，一个敏感的，全面的和早期的生化诊断， AD可通过超灵敏和同时检测错误折叠的寡聚体来开发 由人脑中最常见的三种蛋白质聚集体（Aβ、Tau和αSyn）组成 在人体生物体液（CSF和血浆）中。我们检测这些低聚物的方法 是利用错误折叠的低聚物的功能特性， 单体蛋白的聚合作为检测它们的一种方式。为此，我们发明了 蛋白质错误折叠循环扩增（PMCA）是一种平台技术， 检测到非常少量的与以下相关的有播种能力的错误折叠寡聚蛋白质： 各种蛋白质错误折叠疾病。目前，PMCA已适用于检测错误折叠的朊病毒 在各种生物液体中与朊病毒疾病有关的蛋白质，包括血液和尿液等 在AD和PD患者的CSF中分别存在最近可溶的Aβ和αSyn寡聚体。主要 该项目的目标是使PMCA技术适用于特异性和高灵敏度的检测， 人血液和CSF中错误折叠的Aβ、Tau和αSyn寡聚体，以获得指纹图谱 能够诊断AD并与具有类似临床症状的其他疾病区分开的特征 演示文稿.在这个项目中，我们将使用大的 样本数量，并评估PMCA用于监测疾病进展和 临床前鉴定人的方式发展AD。本项目产生的成果 可能导致开发一种全面的AD生化测试， 只是为了帮助诊断疾病，并将其与其他具有类似特征的疾病区分开来。 临床表现，但也确定人们的方式，以发展AD病理之前， 出现实质性脑损伤和疾病的临床症状。